银屑病患者肠道菌群多样性与表型分析

目的利用新一代16S rDNA全长测序技术比较银屑病患者与健康人的肠道菌群多样性和细菌表型差异。方法纳入12例银屑病患者,以12例正常人作为健康对照,测序分析粪便样品中细菌群落的结构组成和表型特征。结果银屑病患者肠道菌群的多样性指数较健康人显著降低(P0.05),肠道菌群的结构组成明显区别于健康人。在菌门水平上,Firmicutes和Proteobacteria分别在对照组和患者组中高度富集(P0.05)。从菌科到菌种水平上,6个菌科、8个菌属以及6个菌种的相对丰度在患者组和对照组之间存在显著差异(P0.05)。细菌表型的分析结果显示,患者组中厌氧型细菌的相对丰度显著降低(P0.01),兼性厌氧型细菌的相对丰度显著升高(P0.05),细菌对氧化胁迫的耐受性以及细菌生物膜的形成能力增强(P0.05)。结论银屑病患者肠道菌群的多样性较健康人显著降低,其菌落结构和多种细菌表型都发生了明显改变,肠道菌群失调可能是导致银屑病发病的重要原因之一。     

培土清心方对特应性皮炎儿童肠道菌群的影响

目的通过高通量测序手段检测比较特应性皮炎(AD)患儿与健康对照儿童肠道菌群结构差异,并对比培土清心方干预前后对AD患儿的临床疗效及肠道菌群的改变情况,初步探索培土清心方对肠道菌群的影响。方法共纳入60例受试者(实验组、健康对照组各30例),实验组分别在口服培土清心方0周(AD.0W)、4周(AD.4W)进行SCORAD评分并采集粪便样品,对照组在0周时采集粪便样品,统一对粪便样品进行16S rRNA基因测序检测。结果实验组用药4周(AD.4W)SCORD评分降低,差异有统计学意义(P0.05)。肠道菌群测序结果:①Alpha多样性:AD.0W组AD.4W组健康对照组(P0.05)。②差异菌门水平:Firmicutes(厚壁菌门)相对丰度:AD.0W组健康对照组AD.4W组(P0.05);Bacteroidetes(拟杆菌门)相对丰度:AD.4W组健康对照组AD.0W组(P0.05);Faecalibacterium(柔嫩梭菌属)相对丰度:AD.0W组AD.4W组健康对照组,差异有统计学意义(P=0.000.05)。结论特应性皮炎儿童和健康儿童肠道菌群多样性方面无差异,柔嫩梭菌属为健康人优势菌属;培土清心方可改善AD患儿病情,对肠道菌群多样性无明显影响,其可能通过上调厚壁菌门、柔嫩梭菌属,下调拟杆菌门丰度来达到减轻肠道炎症、改善AD的作用。     

高通量测序研究儿童特应性皮炎患者肠道菌群生物多样性特点

目的 探讨特应性皮炎(Atopic dermatitis,AD)儿童患者肠道菌群生物多样性特点。方法 根据纳入标准选取35例AD儿童患者为AD组,27例健康儿童作为对照组(N组)。应用Illumina Miseq测序平台对研究对象粪便细菌的16SrRNA基因V3～V4区进行高通量测序,将测序结果进行肠道菌群差异分析。结果 物种分析显示:在N组中,厚壁菌门、拟杆菌门、变形菌门、放线菌门、其他(低丰度物种),分别占到51.79%、39.06%、5.83%、2.43%和0.90%;在AD组中,厚壁菌门、拟杆菌门、变形菌门、放线菌门、其他(低丰度物种),分别占到58.36%、29.16%、7.47%、4.39%和0.62%。Pan/Core物种分析显示:AD组较N组总物种丰富度低、核心物种数少。多样性指数比较显示:AD组较N组Shannon数值降低,Simpson数值升高,但二者差异均无统计学意义(P0.05)。结论 AD患儿肠道菌群物种丰度及生物多样性与正常儿童存在差异。     

特应性皮炎患儿与正常儿童肠道菌群差异性研究

【摘要】 目的 探讨特应性皮炎（AD） 患儿与正常儿童肠道菌群的差异。方法 收集2015年4月至2017年4月在上海市嘉定区中医医院皮肤科门诊就诊的AD患儿35例,以27例健康儿童作为对照组。取受试者粪便,提取总DNA后,PCR扩增细菌的16SrRNA基因V3 ~ V4区,应用Illumina Miseq测序平台行高通量测序,分析菌群丰度差异。选择两组丰度排名前15的门、属、种分别比较物种差异,采用Wilcoxon秩和检验。结果 两组肠道菌群主要由厚壁菌门、拟杆菌门、变形菌门、放线菌门组成。在菌门水平,AD组与健康对照组拟杆菌门丰度分别为29.16% ± 19.96%、39.06% ± 15.98%（P = 0.042）,梭杆菌门分别为0.06% ± 0.17%、0.50% ± 1.71%（P = 0.041）;在菌属水平,两组拟杆菌属丰度分别为23.77% ± 18.08%、33.1% ± 15.75%（P = 0.029）;在菌种水平,丰度排名前15的菌种两组间分布差异均无统计学意义（均P > 0.05）。结论 特应性皮炎患儿与正常儿童肠道菌群构成及菌群相对丰度存在一定差异。     

寻常痤疮患者肠道-宿主共代谢产物分析

目的分析寻常痤疮患者与健康对照肠道-宿主共代谢产物差异,为肠道菌群参与寻常痤疮发病机制的研究提供理论依据。方法采集43例未经治疗的寻常痤疮患者和43例年龄、性别匹配的健康对照组的粪便标本,对入组者病程、身高、体重、睡眠质量等一般资料进行调查。采用Agilent7890B型气相色谱仪对118种代谢产物进行定量分析,非参数检验比较两组代谢产物差异。结果寻常痤疮组与健康对照组代谢产物组成存在差异;不同病情严重程度痤疮患者代谢产物组成差异不明显;非参数检验分析发现24种肠道-宿主共代谢物含量在痤疮组和对照组之间存在明显的差异(P0.05);另外,寻常痤疮患者睡眠质量较健康对照组差异有统计学意义(P0.05);Pearson相关性分析提示肠道代谢产物与睡眠质量呈明显负相关。结论寻常痤疮患者与健康对照肠道-宿主共代谢产物差异明显,可能成为解释肠道菌群参与寻常痤疮发病机制的纽带。     

基于16S rRNA基因测序的寻常型银屑病患者肠道菌群差异研究

【摘要】 目的 研究成年寻常型银屑病患者和健康人群之间肠道微生物的差异。方法 采集2017年9月至2018年2月于中国医学科学院皮肤病医院确诊的22例寻常型银屑病患者和23例健康对照者粪便样本，提取肠道菌群总DNA扩增，并采用二代16S rRNA基因靶向测序技术测序，分析菌群多样性及菌群分布。对照Silva数据库进行物种注释及分类，采用秩和检验分析各层级样本物种差异；采用QIIME软件（Version 1.9.1）计算OTU数目及α多样性主要指数（Chao1指数、Shannon指数和Simpson指数），t检验分析指数差异；PCoA分析反映样本β多样性差异，置换多元方差分析两组群落组成结构差异；秩和检验及LEfSe分析评估物种差异。结果 银屑病组OTU数目（147.55 ± 57.07）与健康对照组（148.96 ± 50.45）比较，差异无统计学意义（t = 0.088，P = 0.930）。银屑病组Shannon指数（4.08 ± 0.80）、Chao1指数（169.52 ± 63.17）、Simpson指数（0.87 ± 0.07）与健康对照组（分别为4.11 ± 0.94、175.36 ± 53.59、0.86 ± 0.90）比较，差异均无统计学意义（t = 0.12、0.34、0.27，均P > 0.05）。PCoA分析银屑病组与健康对照组的第一主成分差异为49.8%，第二主成分差异为15.62%，置换多元方差分析显示两组间β多样性差异有统计学意义（P = 0.011）。银屑病组与健康对照组有差异的菌群包括在银屑病组中显著升高的厚壁菌门、梭菌纲、梭菌目，丹毒丝菌目、丹毒丝菌科，健康对照组中显著升高的艾普西隆变形杆菌纲、弯曲杆菌目、弯曲杆菌科、弯曲杆菌属，拟杆菌目、拟杆菌科。结论 寻常型银屑病患者与健康人的肠道菌群存在一定差异，肠道菌群有可能成为寻常型银屑病的潜在生物标志物。     

基于16S rRNA基因测序的慢性自发性荨麻疹患者肠道菌群差异研究

【摘要】 目的 研究慢性自发性荨麻疹（CSU）患者和健康人的肠道菌群差异。方法 收集2019年1 - 12月在天津市第一中心医院皮肤科就诊的18例CSU患者（CSU组）及年龄、性别相匹配的18例健康对照（HC组）的粪便标本,提取总DNA,利用16S rRNA测序技术鉴定微生物种类,用生物信息学方法分析菌群差异。采用SPSS 23.0软件对试验数据进行统计学分析。结果 在α多样性方面,CSU组Observed OTU指数161.28 ± 35.47,Chao1指数161.31 ± 35.51,Shannon指数5.15 ± 0.47,Simpson指数0.94 ± 0.03,HC组分别为154.89 ± 54.46、154.92 ± 54.43、4.92 ± 0.88、0.91 ± 0.08,两组差异均无统计学意义（t = 0.417、0.417、0.952、1.116,均P > 0.05）。在β多样性方面,主成分分析显示,两组的第一主成分解释度为6.66%,第二主成分解释度为4.93%,两组菌群结构差异无统计学意义（P = 0.672）。CSU组、HC组肠道菌群Holdemania菌属相对丰度分别为0.04%、0.01%,两组间差异有统计学意义（P = 0.025）。结论 CSU患者与健康人肠道菌群存在一定差异。     

特应性皮炎患儿与健康婴儿肠道菌群的差异性分析

目的:明确特应性皮炎患儿(AD)与健康婴儿肠道菌群的差异性。方法:收集特应性皮炎患儿(AD组)和健康婴儿(对照组)受试者粪便，提取粪便样本总DNA进行定量分析和PCR扩增，采用PacBio Sequel测序仪进行16S rRNA测序。经Wilcoxon秩和检验，比较两组受试者肠道菌群在属、种水平构成性的差异。α多样性和β多样性分析菌群特征，LEfSe分析描述两组菌群代表性差异菌种。结果:共33例特应性皮炎患儿(AD组)和30例健康婴儿(对照组)纳入为研究对象。AD组的菌群物种丰度和均匀度指数较对照组偏低，虽无统计学意义(P>0.05),但两组肠道菌群在组成结构上有显著性差异(P=0.032);采用LEfSe分析方法发现在属和种水平上，AD组弗氏柠檬酸杆菌、产酸克雷伯菌较对照组显著性增加，对照组链球菌属、韦荣氏球菌属、巨球形菌属、唾液乳酸杆菌、第三梭状芽胞杆菌、无害芽胞梭菌等较AD组有显著性增加。结论:AD婴儿肠道菌群相对丰度及生物多样性与正常婴儿存在差异性。     

基于16S rDNA序列分析天疱疮患者皮肤菌群的变化

【摘要】 目的 采用16S rDNA测序技术研究寻常型天疱疮（PV）患者皮肤菌群多样性。方法 收集杭州市第三人民医院皮肤科10例PV患者及10例健康对照,取皮损旁（PV组）及对侧无皮损处皮肤菌群样本（PVn组）,正常对照组取相应部位菌群样本。采用16S rDNA扩增子测序技术,对所有样本的菌群基因进行测序与分类,通过Usearch软件对数据聚类分析,得到可操作分类单元（OTU）,获得门、纲、目、科、属水平的物种丰度。以Observed species指数、Shannon 指数和Simpson指数等反映α多样性,使用主坐标分析法（PCoA）分析β多样性。采用线性判别分析效应量（LEfSe）分析比较各组的差异物种。利用PICRUSt软件进行基因功能预测。2组非参数检验采用Wilcoxon秩和检验,3组间非参数检验采用Kruskal Waills检验。结果 注释到门、纲、目、科、属的OTU分别有2 002、1 869、1 751、1 611、1 120个,3组均以厚壁菌门、放线菌门、拟杆菌门和变形菌门为主,在属水平上,PV组及PVn组葡萄球菌属丰度最高,正常对照组为棒状杆菌属。α多样性分析显示,3组间Observed species指数、Shannon指数、Simpson指数差异均有统计学意义（均P < 0.05）,PV组Shannon指数（3.24 ± 1.30）和Simpson指数（0.70 ± 0.19）均低于正常对照组（P < 0.05）。PCoA分析显示,3组间β多样性差异无统计学意义（P = 0.054）。秩和检验显示,3组间丰度差异有统计学意义的物种共32个,其中相对丰度较高的有富集于PV组的芽孢杆菌纲及富集于正常对照组的微球菌属、短波单胞菌属等。按病程 < 3个月、 ≥ 3个月分组,PV长病程组中富集的物种有梭状芽孢杆菌目、颤杆菌属及鞘氨醇单胞菌属等;短病程组γ变形菌纲富集。功能预测显示,与感染性疾病相关的基因功能富集于天疱疮组。结论 16S rDNA测序技术应用于天疱疮微生物组学研究,证实PV皮肤菌群多样性及组成结构与正常人存在一定差异。     

儿童特应性皮炎患者肠道菌群中益生菌的构成比研究

目的：比较特应性皮炎(AD)患儿与正常儿童肠道菌群中益生菌的构成差异。方法：应用Illumina MiSeq测序平台对特应性皮炎患儿和健康对照儿童粪便细菌的16SrRNA基因V3-V4区进行高通量测序,比较两组益生菌属、种水平的构成差异。结果: 共纳入35例 AD 患儿和27例正常对照,在属水平,AD组患者双歧杆菌属(Bifidobacterium)物种丰度较对照组升高,而乳酸杆菌属(Lactobacillus)物种丰度较对照组降低,但二者差异均无统计学意义（P＞0.05）。在种水平,AD组患者双歧杆菌属下Bifidobacterium pseudocatenulatum DSM 20438 JCM 1200 LMG 10505、unclassified_g__Bifidobacterium、Bifidobacterium_adolescentis、Bifidobacterium_animalis 物种丰度较对照组升高,而Bifidobacterium_breve物种丰度降低,差异均无统计学意义（P＞0.05）;AD组患者乳酸杆菌属下Lactobacillus_salivarius物种丰度较对照组降低,差异无统计学意义（P＞0.05）,而unclassified_g__Lactobacillus、Lactobacillus_plantarum_subsp._plantarum物种丰度升高,差异有统计学意义（P＜0.05）。结论:特应性皮炎患儿肠道菌群中益生菌物种丰度与正常儿童存在差异。     

Skin microbiome differences relate to the grade of acne vulgaris

The skin microbiome plays important roles in the pathogenesis and development of acne. We aimed to investigate the facial skin microbiome of acne and microbiome differences related to different grades of acne. Skin swabs from nine healthy controls and 67 acne patients were collected, and the skin microbiomes were analyzed using 16S rRNA gene sequencing. Compared with healthy controls, acne patients harbored significantly altered skin microbiomes. The skin microbiomes of patients with grade 1–3 acne were similar, but patients with grade 4 acne showed a significantly different skin microbiome compared with grade 1–3 acne, including increased alpha diversity and increased proportions of four Gram‐negative bacteria (Faecalibacterium, Klebsiella, Odoribacter and Bacteroides). In conclusion, acne patients harbored an altered skin microbiome, and more significant dysbiosis was found in patients with grade 4 acne (severe acne). Our findings may provide evidence for the pathogenic mechanisms of acne and microbial‐based strategies to avoid and treat acne, especially grade 4 acne.     

酒渣鼻患者鼻部皮肤蠕形螨寄生对局部微生态的影响

【摘要】 目的 分析酒渣鼻患者鼻部蠕形螨寄生与鼻部皮肤微生物群落的关系。方法 2017年5月至2019年6月于佛山市顺德区慢性病防治中心皮肤科收集酒渣鼻患者与面部健康对照者各14例，酒渣鼻患者中早期8例，中期6例。采集受试者鼻翼和鼻唇沟皮肤微生物样品，提取DNA，采用宏基因组测序和生物信息学分析。以蠕形螨及微生物reads数的构成比反映菌种相对丰度。计算Shannon指数评估微生物α多样性。分析基于菌种相对丰度的主成分（PCA）以评估β多样性。计数资料的比较采用两独立样本t检验，蠕形螨与微生物含量间关系采用Pearson相关性分析。结果 酒渣鼻组鼻部皮肤蠕形螨相对含量（1.647% ± 0.389%）高于健康组（0.448% ± 0.089%，t = 2.92，P = 0.007）。蠕形螨的相对含量与细菌相对含量呈负相关（r = -0.95，P < 0.001），与真菌相对含量呈正相关（r = 0.76，P < 0.001）。酒渣鼻组鼻细菌、真菌群落Shannon指数（0.91 ± 0.17、1.261 ± 0.045）显著高于健康组（0.47 ± 0.12、0.549 ± 0.071，t = 2.17 、8.48，P < 0.05）；两组的主成分分析结果示，仅细菌群落显著不同（t = 2.32，P = 0.029），而真菌群落无差异（t = 0.82，P = 0.461）。此外，中期酒渣鼻患者蠕形螨相对含量显著高于早期（t = 6.56，P < 0.001）；早、中期患者中细菌和真菌的Shannon指数差异无统计学意义（均P > 0.05），主成分分析结果示细菌和真菌的群落结构差异均有统计学意义（均P < 0.05）。结论 蠕形螨在鼻部皮肤的寄生可能影响鼻部皮肤微生态群落结构。     

Gut microbiota alterations in moderate to severe acne vulgaris patients

Acne vulgaris is a chronic inflammatory dermatosis affecting approximately 85% of adolescents. There are many factors contributing to the development of this ailment. A recent study indicated that gut microbiota takes part in the pathogenesis of acne. We aimed to investigate the link between acne vulgaris and gut microbiota. A total of 31 moderate to severe acne vulgaris patients and 31 healthy controls were enrolled. We collected their feces, and gut microbiota was evaluated by the hypervariable regions of 16S rRNA genes through high‐throughput sequencing. We identified links between acne vulgaris and changes of gut microbiota. At the phylum level, Actinobacteria (0.89% in acne patients and 2.84% in normal controls, P = 0.004) was decreased and Proteobacteria (8.35% in acne patients and 7.01% in normal controls, P = 0.031) was increased. At the genus level, Bifidobacterium, Butyricicoccus, Coprobacillus, Lactobacillus and Allobaculum were all decreased. The observed difference in genera between acne patients and healthy controls provides a new insight into the link between gut microbiota changes and acne vulgaris risk.     

Isotretinoin and lymecycline treatments modify the skin microbiota in acne

Oral retinoids and tetracyclines have a major role in acne treatment. Here, we report for the first time the effect of isotretinoin and lymecycline therapy on the skin microbiota in cheek, back and armpit swab samples of acne vulgaris patients using 16S ribosomal RNA (16S rRNA) gene amplicon sequencing. Propionibacterium acnes was the most common in sebaceous areas of healthy and untreated acne skin and more abundant in back than cheek samples. Five taxa, including a Streptococcus taxon, differed significantly between the cheek samples of healthy controls and acne patients, and acne severity was positively correlated with the abundance of Propionibacterium. Both treatments reduced clinical acne grades and the abundance of Propionibacterium, while the abundance of several other taxa was significantly higher in treated cheek samples compared with untreated ones. Less variation was observed in back samples and none in armpit samples. There were no differences in alpha diversity between control and acne patients in any of the sampled skin areas, but the diversity of the microbiota on the cheek and the back was significantly increased after acne treatments. This study provides insight into the skin microbiota in acne and how it is modulated by systemic acne treatment.     

Gut microbiota dysbiosis

Human beings and other animals live with a complex mixture of different bacteria (called microbiota) on the skin and in the gut, and these micro-organisms play an important role in helping us to remain healthy. Alterations in the proportions of different bacteria are found in several disease states; this is called dysbiosis. The mixture of different bacteria in the healthy gut plays an important role in developing and maintaining a healthy immune system. Although patients with psoriasis are known to have altered microbiota on their skin, compared with unaffected individuals, little is known about the proportions of different bacteria in their guts. The authors, working in several different departments in Spain, suggest that changes in the gut microbiota may play a part in the development of psoriasis. They compared the gut microbiota in 19 patients with psoriasis with 20 healthy controls (people without psoriasis) living in the same region, by extracting bacterial DNA from fresh samples of faeces. The authors found a reduced diversity of bacteria in the samples from psoriatic patients, with a relative abundance of some bacterial species not found in the healthy controls. They suggest that further studies are needed to compare the patterns of microbiota in patients with different degrees of psoriasis severity, and in different patterns of psoriasis. This could help to establish whether gut dysbiosis plays a role in the development of the disease; if so, there could be a place for therapies aimed at changing the bacterial composition of the gut, such as transplantation of faecal bacteria or the use of probiotic treatments.     

Tissue and blood superoxide dismutase activities and malondialdehyde levels in different clinical severities of acne vulgaris

Background Acne vulgaris is one of the commonest dermatological diseases and its pathogenesis is multifactorial. Objectives To determine the role of oxidative stress in acne vulgaris and to determine a possible link with the clinical severity. Methods Twenty‐three patients with different grades of acne vulgaris and 23 age‐ and sex‐matched controls were enrolled. Superoxide dismutase (SOD) activities and malondialdehyde (MDA) levels were measured spectrophotometrically at tissue and blood levels. Results There were no significant differences in SOD activities and MDA levels between patients and controls. However, significant differences were found in patients with severe acne in comparison with those with mild and moderate acne. Moreover, comparison between different patient subgroups and controls revealed statistically significantly higher SOD activities in patients with mild acne in comparison with patients with moderate and severe acne, and with controls. Furthermore, severe acne showed statistically significantly lower SOD activities and higher MDA levels when compared with other patient subgroups and controls. Conclusions Oxidative stress exists in patients with acne vulgaris and may play a role in aetiopathogenesis and/or progression of the disease. The addition of drugs with antioxidative effects seems to be valuable in the treatment of acne vulgaris.